Identification of myosin-binding sites on the actin sequence.
Identification of myosin-binding sites on the actin sequence.
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DOI：
10.1021/bi00258a020
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发表时间：
1982-07
期刊：
影响因子：
2.9
通讯作者：
K. Sutoh
中科院分区：
文献类型：
--
作者：
K. Sutoh
肌动蛋白和胰蛋白酶处理的肌球蛋白亚片段 1 (S1) 的严格复合物，其重链被切割成三个片段（20K、25K 和 50K），并与零长度交联剂 1-乙基-3-[3-(二甲基氨基)丙基]碳二亚胺进行交联。交联反应生成三种类型的交联产物，表观分子量分别为65K、68K和95K。 65K、68K和95K产物分别是S1重链肌动蛋白-20K片段、肌动蛋白-碱性轻链1和S1重链肌动蛋白-50K片段的共价连接复合物。肌动蛋白序列上 S1 重链和轻链的交联位点已通过用溴化氰或羟胺消化交联产物，然后在十二烷基硫酸钠凝胶上绘制所得肽来确定。结果表明，肌动蛋白N端部分1、2、3、4和11位酸性残基是S1重链20K和50K片段的交联位点，而其C端部分360、362和363位酸性残基是碱性轻链1的交联位点。
The rigor complex of actin and trypsin-treated myosin subfragment 1 (S1) whose heavy chain was cleaved into three fragments (20K, 25K, and 50K) was cross-linked with a zero-length cross-linker, 1-ethyl-3-[3-(dimethyl-amino) propyl]carbodiimide. The cross-linking reaction generated three types of cross-linked products with apparent molecular weights of 65K, 68K, and 95K. The 65K, 68K, and 95K products were covalently linked complexes of actin-20K fragment of the S1 heavy chain, actin-alkaline light chain 1, and actin-50K fragment of the S1 heavy chain, respectively. Cross-linking sites of S1 heavy and light chains on the actin sequence have been determined by digesting the cross-linked products with cyanogen bromide or with hydroxylamine and then mapping resulting peptides on sodium dodecyl sulfate gels. The result indicates that some of the N-terminal acidic residues of actin at positions 1, 2, 3, 4, and 11 are cross-linking sites of the 20K and 50K fragments of the S1 heavy chain while some of its C-terminal acidic residues at positions 360, 362, and 363 are cross-linking sites of the alkaline light chain 1.